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The purpose of this study was to examine the possibility of separation and quantification of serum alkaline phosphatase (ALP) isozymes in rats by wheatgerm lectin affinity electrophoresis. Cellulose acetate electrophoresis of the liver and bone ALPs without lectin results in overlapping bands, but in the presence of lectin, the mobility of the band of bone enzyme was retarded and well separated from the liver enzyme band. With this affinity electrophoretic method, we determined the serum ALP isozymes in fed and fasting rats grouped by age. As a result, the absolute activity of bone isozyme showed a downward trend with age in the fed and fasting rats. The serum ALP activity was steadily higher in fed rats than in fasting rats, and the increase was due to intestinal ALP isozyme. There was low activity bordering complete absence in liver isozyme under both nutritional conditions. The affinity electrophoretic method provided a rapid, reproducible, and relatively simple technique for further clinical characterization of ALP isozyme in the rat serum. 相似文献
4.
Root elongation of Alaska pea seedling was suppressed by higherconcentrations of growth retardants, CCC and ancymidol, thanthose required for shoot elongation. Gibberellic acid (GA3)led to recovery of ancymidol-inhibited elongation, with theconcentration (1 nM) required for roots being lower than thatfor shoots (10 µM). Ancymidol caused swelling of corticalcells in the elongating zone of the root, while GA3 completelycanceled this. These results suggest that roots require muchless gibberellin than shoots for normal elongation growth. Growth kinetics recorded by a computer-regulated rhizometerindicated that the lag periods for growth suppression by ancymidoland growth recovery by GA3 were about 10 h and 7 h, respectively. The composition of the cell wall sugars changed remarkably alongthe root axis from the tip to the base. The arabinose contentwas highest in the tip and rapidly decreased toward the base,whereas galactose complementarily increased toward the base.The thickened zone of ancymidol-treated roots had a higher galactosecontent than GA3-treated slender roots. Other neutral sugarswere not significantly influenced by ancymidol and/or GA3. Theseresults suggest that ancymidol makes cells short and thick withgalactose-rich cell walls while GA3 keeps cells extensible andslender with galactose-poor cell walls. (Received March 3, 1987; Accepted December 4, 1987) 相似文献
5.
From dilution series in defined mineral medium, a marine iregular coccoid methanogenic bacterium (strain MTP4) was isolated
that was able to grow on methanethiol as sole source of energy. The strain also grew on dimethylsulfide, mono-, di-, and trimethylamine,
methanol and acetate. On formate the organism produced methane without significant growth. Optimal growth on MT, with doubling
times of about 20 h, occurred at 30°C in marine medium. The isolate required p-aminobenzoate and a further not identified
vitamin. Strain MTP4 had a high tolerance to hydrogen sulfide but was very sensitive to mechanical forces or addition of detergents
such as Triton X-100 or sodium dodecylsulfate. Methanethiol was fermented by strain MTP4 according to the following equation:
相似文献
6.
High-affinity binding of3H-folate in Triton X-100 solubilized membranes of human liver displayed characteristics, e.g. apparent positive cooperativity, which are typical of specific folate binding. Ultrogel® AcA 44 chromatography of solubilized membranes saturated with3H-folate revealed a major peak of 100 kDa and a minor peak of 25 kDa. The 100 kDa peak could represent a hydrophobic membrane associated molecular form of the protein. This notion was supported by the fact that the two peaks had identical molecular weights as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis with immunoblotting. 相似文献
7.
M Nakano K Kasai K Yoshida T Tanimoto Y Tamaki T Tobita 《Journal of biochemistry》1989,105(1):133-137
1. CD spectra showed that the fowl protamine, galline, has an unordered structure rich in reverse turns in neutral solution. Eight reverse turns were predicted to be present in the galline molecule on the basis of its amino acid sequence. Spectrophotometric analyses revealed that galline efficiently bound to DNA in 0.25 mM EDTA/10 mM Tricine-HCl, pH 7.4, but hardly so in 30 mM NaCl/3 mM sodium citrate, pH 7.0. Citrate ions bound specifically to the galline molecule, causing a conformational change in it. As a result, galline could not interact with DNA. 2. The concentration of unbound galline in a mixture of DNA and galline in 100 mM NaCl/50 mM Tricine-HCl, pH 7.4, at 37 C was determined by measurement of the intrinsic fluorescence of tyrosine residues of galline in the supernatant after ultracentrifugation of the mixture. The Scatchard plot showed positive co-operativity in the binding of galline to DNA and the binding parameters were determined: the co-operative binding constant (Kc) = 3.3 X 10(7)M-1, the co-operativity factor (q) = 800, and the number of nucleotides of DNA occupied by one galline molecule (n) = 28. The Kc and q values were intermediate between those for clupeine Z from herring sperm and S-methyl protamine from boar sperm. That is, the binding constants of protamine as to DNA decrease in the order of herring, fowl, and boar, while the co-operativities in binding increase in that order. 相似文献
8.
Anaerobic degradation of methylmercaptan and dimethyl sulfide by newly isolated thermophilic sulfate-reducing bacteria. 总被引:5,自引:4,他引:1
![]() The complete oxidation of methylmercaptan (MSH) and dimethyl sulfide (DMS) with sulfate or nitrate as electron acceptors was observed in enrichment cultures and dilution series using thermophilic fermentor sludge as the inoculum. Three new strains of thermophilic sulfate reducers were isolated in pure culture (strains MTS5, TDS2, and SDN4). Strain MTS5 grew on MSH and strain TDS2 grew on DMS whereas strain SDN4 grew on either MSH or DMS. The cellular growth yields were 2.57 g (dry weight)/mol of MSH for strain MTS5 and 6.02 g (dry weight)/mol of DMS for strain TDS2. All strains used sulfate, sulfite, or thiosulfate as electron acceptors, but only strain SDN4 used nitrate. DMS and MSH were oxidized to CO2 and sulfide with either sulfate or nitrate as the electron acceptor. Sulfate was stoichiometrically reduced to sulfide while nitrate was reduced to ammonium. All strains were motile rods, required biotin for growth, lacked desulfoviridin, had DNA with G+C contents of 48 to 57 mol% and probably belonged to the genus Desulfotomaculum. This is the first report of the oxidation of MSH and DMS by pure cultures of sulfate-reducing bacteria. 相似文献
9.
High-affinity folate binding in human prostate 总被引:1,自引:0,他引:1
Binding of3H-folate in Triton X-100 solubilized human prostate homogenate was of a high-affinity type and displayed apparent positive cooperativity typical of specific folate binding. Radioligand dissociation was slow at pH 7.4, but rapid at pH 3.5. Gel chromatography reveled two major folate binding proteins (Mr100 and 25kDa), but only one single band (Mr65–70 kDa) was detectable on SDS-PAGE and immunoblotting with rabbit-anti human milk folate binding protein. Concentration of folate binding protein in prostate homogenate expressed as maximum3H-folate binding was 1.10 nmol/g protein, and the cross-reactivity with rabbit-anti human milk folate binding protein serum was 15% as determined by an enzyme-linked immunosorbent assay (median values; n=6). 相似文献
10.
High-affinity3H-folate binding in Triton X-100 solubilized human mammary gland tissue displayed characteristics, e.g. apparent positive cooperativity and increasing affinity with decreasing concentration of folate binding protein, shown to be typical of specific folate binding. Radioligand dissociation was slow at pH 7.4. A major fraction of the bound radioligand dissociated rapidly at pH 3.5, while a residual binding of 20% persisted even after prolonged dialysis at pH 3.5. Gel chromatography revealed two major folate binding proteins (Mr100 kDa and 25 kDa). However, only one single band was detectable on SDS-PAGE immunoblotting. The highest folate binding activity per g protein was associated with the upper triglyceride-containing layer of the 1000 g supernatant of the homogenate. The folate binding protein extracted from this layer had a low cross-reactivity (<5%) with rabbit antibodies against 25 kDa human milk folate binding protein. The folate binding protein in the 1000 g pellet and the aqueous phase of the 1000 g supernatant was present at a low concentration and had a cross-reactivity of 100%. 相似文献
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